U.S. patent application number 14/289450 was filed with the patent office on 2014-11-27 for co-administration of pimavanserin with other agents.
This patent application is currently assigned to Acadia Pharmaceuticals Inc.. The applicant listed for this patent is Acadia Pharmaceuticals Inc.. Invention is credited to Uli HACKSELL, Krista MCFARLAND.
Application Number | 20140349976 14/289450 |
Document ID | / |
Family ID | 40350231 |
Filed Date | 2014-11-27 |
United States Patent
Application |
20140349976 |
Kind Code |
A1 |
HACKSELL; Uli ; et
al. |
November 27, 2014 |
CO-ADMINISTRATION OF PIMAVANSERIN WITH OTHER AGENTS
Abstract
As disclosed herein, co-administration of pimavanserin with an
agent that ameliorates one or more cholinergic abnormalities can
have a synergistic effect on the efficacy of the agent. Disclosed
herein are compositions which include pimavanserin in combination
with an agent that ameliorates one or more cholinergic
abnormalities. Also disclosed herein are methods for ameliorating
or treating a disease condition characterized by one or more
cholinergic abnormalities that can include administering
pimavanserin in combination with an agent that ameliorates one or
more cholinergic abnormalities.
Inventors: |
HACKSELL; Uli; (San Diego,
CA) ; MCFARLAND; Krista; (San Marcos, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Acadia Pharmaceuticals Inc. |
San Diego |
CA |
US |
|
|
Assignee: |
Acadia Pharmaceuticals Inc.
San Diego
CA
|
Family ID: |
40350231 |
Appl. No.: |
14/289450 |
Filed: |
May 28, 2014 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
12234573 |
Sep 19, 2008 |
|
|
|
14289450 |
|
|
|
|
60974426 |
Sep 21, 2007 |
|
|
|
60986250 |
Nov 7, 2007 |
|
|
|
61050976 |
May 6, 2008 |
|
|
|
Current U.S.
Class: |
514/129 ;
514/139; 514/292; 514/297; 514/329 |
Current CPC
Class: |
A61P 25/28 20180101;
A61K 31/4465 20130101; A61P 25/18 20180101; A61P 25/08 20180101;
A61P 25/32 20180101; A61K 31/4468 20130101; A61P 25/30 20180101;
A61P 43/00 20180101; A61P 25/24 20180101; A61P 25/22 20180101; A61K
45/06 20130101; A61K 31/473 20130101; A61P 25/04 20180101; A61P
25/00 20180101; A61P 25/20 20180101; A61P 25/16 20180101; A61P
25/14 20180101 |
Class at
Publication: |
514/129 ;
514/329; 514/297; 514/292; 514/139 |
International
Class: |
A61K 31/4465 20060101
A61K031/4465; A61K 45/06 20060101 A61K045/06; A61K 31/473 20060101
A61K031/473 |
Claims
1-32. (canceled)
33. A method for ameliorating or treating a disease condition
characterized by one or more cholinergic abnormalities, comprising
administering to a subject with the disease condition: (a)
pimavanserin, or a pharmaceutically acceptable salt thereof; and
(b) a cholinesterase inhibitor, wherein the cholinesterase
inhibitor is metrifonate, physostigmine, neostigmine,
pyridostigmine, demarcarium, rivastigmine, aldicarb, bendiocarb,
bufencarb, carbaryl, carbendazim, carbetamide, carbofuran,
chlorbufam, chloropropham, ethiofencarb, formetanate, methiocarb,
methomyl, oxamyl, phenmedipham, pinmicarb, pirimcarb, propamocarb,
propham, propoxus, donepezil, tacrine, edrophonium, echothiophate,
diisopropyl fluorophosphate, dimebon, Huperzine A,
((2-[2-(1-benzylpiperidin-4-yl)ethyl]-2,3-dihydro-9-methoxy-1H-pyrrolo[3,-
4-b]quinolin-1-one hemifumarate)), zanapezil, phenserine,
quilostigmine, ganstigmine, butyrophenones, imipramines, tropates,
phencyclidines, curariforms, ethephon, ethopropazine, iso-OMPA,
tetrahydrofurobenzofuran cymserine, N.sup.1-phenethyl-norcymserine,
N.sup.8-benzylnorcymserine, N.sup.1,N.sup.8-bisnorcymserine,
N.sup.1--N.sup.8-bisbenzylnorphysostigmine,
N.sup.1,N.sup.8-bisbenzylnorphenserine, or
N.sup.1,N.sup.8-bisbenzylnorcymserine.
34. The method of claim 33, wherein the cholinesterase inhibitor is
metrifonate, pyridostigmine, rivastigmine, bufencarb, carbaryl,
carbofuran, chlorbufam, chloropropham, ethiofencarb, formetanate,
methomyl, oxamyl, pinmicarb, donepezil, tacrine, edrophonium,
diisopropyl fluorophosphate, dimebon, zanapezil, phenserine,
quilostigmine, phencyclidines, curariforms, ethopropazine,
tetrahydrofurobenzofuran cymserine, N.sup.1-phenethyl-norcymserine,
N.sup.8-benzylnorcymserine, or N.sup.1,N.sup.8-bisnorcymserine.
35. The method of claim 33, wherein the cholinesterase inhibitor is
metrifonate, rivastigmine, dimebon, phencyclidines, diisopropyl
fluorophosphate, or donepezil.
36. The method of claim 33, wherein the cholinesterase inhibitor is
metrifonate.
37. The method of claim 33, wherein the cholinesterase inhibitor is
rivastigmine.
38. The method of claim 33, wherein the cholinesterase inhibitor is
dimebon.
39. The method of claim 33, wherein the cholinesterase inhibitor is
diisopropyl fluorophosphate.
40. The method of claim 33, wherein the cholinesterase inhibitor is
donepezil.
41. The method of claim 33, wherein a pharmaceutically acceptable
salt of pimavanserin is administered, wherein the pharmaceutically
acceptable salt is a tartrate or a hemi-tartrate salt.
42. The method of claim 41, wherein the pharmaceutically acceptable
salt of pimavanserin is a crystalline Form C tartrate salt.
43. The method of claim 33, wherein the amount of pimavanserin, or
a pharmaceutically acceptable salt thereof, is from about 0.1 mg to
about 500 mg per day.
44. The method of claim 33, wherein the amount of pimavanserin, or
a pharmaceutically acceptable salt thereof, is from about 1 mg to
about 250 mg per day.
45. The method of claim 33, wherein the amount of pimavanserin, or
a pharmaceutically acceptable salt thereof, is about 1 mg to about
40 mg per day.
46. The method of claim 33, wherein the amount of pimavanserin, or
a pharmaceutically acceptable salt thereof, is about 10 mg per
day.
47. The method of claim 33, wherein the amount of pimavanserin, or
a pharmaceutically acceptable salt thereof, is about 20 mg per
day.
48. The method of claim 33, wherein the amount of pimavanserin, or
a pharmaceutically acceptable salt thereof, is about 40 mg per
day.
49. The method of claim 33, wherein pimavanserin is administered
orally, parenterally, or intravenously.
50. The method of claim 49, wherein pimavanserin is administered
orally.
51. The method of claim 50, wherein pimavanserin is administered as
one or more tablets.
Description
RELATED APPLICATION INFORMATION
[0001] This application claims priority to U.S. Provisional
Application Ser. No. 60/974,426, entitled "N-SUBSTITUTED PIPERIDINE
DERIVATIVES AS SEROTONIN RECEPTOR AGENTS," filed on Sep. 21, 2007;
60/986,250, entitled "CO-ADMINISTRATION OF PIMAVANSERIN WITH OTHER
AGENTS," filed Nov. 7, 2007 and 61/050,976; entitled
"CO-ADMINISTRATION OF PIMAVANSERIN WITH OTHER AGENTS" filed May 6,
2008, all of which are incorporated herein by reference in their
entireties, including all drawings, for all purposes.
BACKGROUND
[0002] 1. Field
[0003] The present application relates to the fields of chemistry
and medicine. More particularly, disclosed herein are methods and
compositions that can be used to treat disease conditions such as
those characterized by cholinergic abnormalities.
[0004] 2. Description of the Related Art
[0005] Conditions associated with cognitive impairment, such as
Alzheimer's disease, are accompanied by loss of acetylcholine in
the brain. This is believed to be the result of degeneration of
cholinergic neurons in the basal forebrain, which widely innervate
multiple areas of the brain, including the association cortices and
hippocampus that are critically involved in higher processes.
[0006] Efforts to increase acetylcholine levels have focused on
increasing levels of choline, the precursor for acetylcholine
synthesis, and on blocking acetylcholinesterase (AChE), the enzyme
that metabolizes acetylcholine. To date, attempts to augment
central cholinergic function through the administration of choline
or phosphatidylcholine have not been successful. AChE inhibitors
have shown therapeutic efficacy, but have been found to have
frequent cholinergic side effects due to excessive increases in
acetylcholine in the periphery-mediated and central-mediated,
including abdominal cramps, nausea, vomiting, and diarrhea. These
gastrointestinal side effects have been observed in about a third
of the patients treated. In addition, some AChE inhibitors, such as
tacrine, have also been found to cause significant hepatotoxicity
with elevated liver transaminases observed in about 30% of
patients. Consequently, the adverse effects of AChE inhibitors have
severely limited their clinical utility.
[0007] Attempts to ameliorate the effects of decreased
acetylcholinergic transmission by direct agonism of the muscarinic
M.sub.1 subtype of acetylcholine receptor has not proven successful
because known muscarinic agonists lack specificity in their actions
at the various muscarinic receptor subtypes, leading to
dose-limiting side effects that limit their clinical utility. For
example, the M.sub.1 muscarinic agonist arecoline has been found to
be an agonist of M.sub.2 as well as M.sub.3 muscarinic receptor
subtypes, and is not very effective in treating cognitive
impairment, most likely because of dose-limiting M.sub.2 and
M.sub.3 receptor mediated side effects. Similarly, xanomeline
(Shannon et al., J. Pharmacol. Exp. Ther. 1994, 269, 271; Shannon
et al., Schizophrenia Res. 2000, 42, 249) is an M.sub.1/M.sub.4
preferring muscarinic receptor agonist shown to reduce psychotic
behavioral symptoms in Alzheimer's disease patients (Bodick et al.,
Arch. Neurol. 1997, 54, 465), which also suffers from treatment
induced side effects that severely limit its clinical utility.
SUMMARY
[0008] One embodiment described herein relates to a composition
that can include: pimavanserin, or a salt, a solvate, a polymorph
or an isolated, substantially pure metabolite thereof; and an agent
that ameliorates one or more cholinergic abnormalities. In some
embodiments, the agent that ameliorates one or more cholinergic
abnormalities can be a cholinesterase inhibitor. In an embodiment,
the cholinesterase inhibitor can be selected from an
acetyicholinesterase inhibitor and a butyrylcholinesterase
inhibitor. Exemplary cholinesterase inhibitors include, but are not
limited to, metrifonate, physostigmine, neostigmine,
pyridostigmine, ambenonium, demarcarium, rivastigmine, aldicarb,
bendiocarb, bufencarb, carbaryl, carbendazim, carbetamide,
carbofuran, chlorbufam, chloropropham, ethiofencarb, formetanate,
methiocarb, methomyl, oxamyl, phenmedipham, pinmicarb, pirimicarb,
propamocarb, propham, propoxur, galantamine, donepezil (E-2020),
tacrine, edrophonium, phenothiazines, echothiophate, diisopropyl
fluorophosphate, dimebon, Huperzine A, T-82
((2-[2-(1-benzylpiperidin-4-yl)ethyl]-2,3-dihydro-9-methoxy-1H-pyrrolo[3,-
4-b]quinolin-1-one hemifumarate)), TAK-147 (zanapezil), phenserine,
quilostigmine, ganstigmine, butyrophenones, imipramines, tropates,
phencyclidines, curariforms, ethephon, ethopropazine, iso-OMPA,
tetrahydrofurobenzofuran cymserine, N.sup.1phenethyl-norcymserine,
N.sup.8-benzylnorcymserine, N.sup.1,N.sup.8-bisnorcymserine,
N.sup.1--N.sup.8-bisbenzylnorphysostigmine,
N.sup.1,N.sup.8-bisbenzylnorphenserine and
N.sup.1,N.sup.8-bisbenzylnorcymserine. In an embodiment, the
cholinesterase inhibitor can be tacrine.
[0009] In other embodiments, the agent that ameliorates one or more
cholinergic abnormalities can be a muscarinic receptor agonist.
Examples of suitable muscarinic receptor agonists include, but are
not limited to, xanomeline, carbamylcholine, oxotremorine,
methacholine, bethanechol, cevimeline (AF102B), AF150(S), AF267B,
aceclidine, arecoline, milameline, talsaclidine, pilocarpine and
(S)-2-ethyl-8-methyl-1-thia-4,8-diaza-spiro[4.5]decan-3-one (Torrey
Pines NGX267). In an embodiment, the muscarinic receptor agonist
can be xanomeline.
[0010] In still other embodiments, the agent that ameliorates one
or more cholinergic abnormalities can be a glutamatergic
antagonist. Suitable glutamatergic antagonists include, but are not
limited to, amantadine, dextromethorphan, dextrorphan, ibogaine,
ketamine, tramadol, methadone, and memantine. In an embodiment, the
glutamatergic antagonist can be memantine.
[0011] In some embodiments, the agent that ameliorates one or more
cholinergic abnormalities can be a cholinergic agonist. Exemplary
cholinergic agonists include, but are not limited to, pramiracetam,
piracetam, oxiracetam, choline-L-alfoscerate, nebracetam,
besipirdine, and taltirelin.
[0012] In other embodiments, the agent that ameliorates one or more
cholinergic abnormalities can be a carnitine acetyltransferase
stimulant. Exemplary carnitine acetyltransferase stimulants
include, but are not limited to, levocarnitine, ST-200
(acetyl-1-carnitine), and nefiracetam.
[0013] In still other embodiments, the agent that ameliorates one
or more cholinergic abnormalities can be an acetylcholine release
stimulant. Exemplary acetylcholine release stimulants include, but
are not limited to, SIB-1553A
((+/-)-4-[[2-(1-methyl-2-pyrrolidinyl)ethyl]thio]phenol
hydrochloride) and T-588
((1R)-1-benzo[b]thiophen-5-yl-2-[2-(diethylamino) ethoxy]ethan-1-ol
hydrochloride).
[0014] In even other embodiments, the agent that ameliorates one or
more cholinergic abnormalities can be a choline uptake stimulant.
In an embodiment, the choline uptake stimulant can be MKC-231
(2-(2-oxopyrrolidin-1-yl)-N-(2,3-dimethyl-5,6,7,8-tetrahydrofuro[2,3-b]qu-
inolin-4-yl)acetoamide).
[0015] In some embodiments, the agent that ameliorates one or more
cholinergic abnormalities can be a nicotinic acetylcholine receptor
agonist. Examples of suitable nicotinic acetylcholine receptor
agonist include, but are not limited to, nicotine, ABT-418,
ABT-089, SIB-1508Y, A-582941, DMXB-A, Sazetidine-A, Varenicline and
TC-1734.
[0016] In some embodiments, the agent that ameliorates one or more
cholinergic abnormalities can be a 5-HT6 antagonist and/or 5-HT6
inverse agonist. Suitable of 5-HT6 antagonists and/or 5-HT6 inverse
agonists include, but are not limited to, SB-742457, SB-271046,
SB-399885, SB-357134, SB-258585, RO-436854, RO-0406790 and
RO-65-7674.
[0017] Another embodiment described herein relates to a method for
ameliorating or treating a disease condition characterized by one
or more cholinergic abnormalities that can include administering a
therapeutically effective amount of one or more compositions
described herein to a subject suffering from the disease condition
characterized by one or more cholinergic abnormalities.
[0018] Still another embodiment described herein relates to a
method for ameliorating or treating a disease condition
characterized by one or more cholinergic abnormalities that can
include administering a therapeutically effective amount of
pimavanserin in combination with a therapeutically effective amount
of an agent that ameliorates one or more cholinergic abnormalities,
such as those described herein, to a subject suffering from the
disease condition characterized by one or more cholinergic
abnormalities. In an embodiment, pimavanserin can be administered
before the agent. In another embodiment, the pimavanserin can be
administered after the agent. In still another embodiment, the
pimavanserin can be administered at approximately the same time as
the agent. Exemplary disease conditions include, but are not
limited to, a neuropsychiatric disorder, a neurodegenerative
disorder and an extrapyrimidal disorder. Other examples of suitable
disease conditions include, but are not limited to, cognitive
impairment, forgetfulness, confusion, memory loss, an attention
deficit disorder, depression, pain, a sleep disorder, psychosis, a
hallucination, aggressiveness, and paranoia. In an embodiment, the
psychosis can be selected from drug-induced psychosis,
treatment-induced psychosis and psychosis associated with a
disease. In some embodiments, the disease the psychosis is
associated with can be dementia, post traumatic stress disorder,
Alzheimer's disease, Parkinson's disease and schizophrenia. In an
embodiment, the psychosis can be Alzheimer's disease-induced
psychosis. In an embodiment, the psychosis can be
schizophrenia-induced psychosis. In an embodiment, the psychosis
can be dementia-related psychosis. Still other exemplary disease
conditions include but are not limited to, a neurodegenerative
disease, Alzheimer's disease, Parkinson's disease, Huntington's
chorea, Friederich's ataxia, Gilles de la Tourette's syndrome, Down
Syndrome, Pick disease, dementia, clinical depression, age-related
cognitive decline, attention-deficit disorder, sudden infant death
syndrome, and glaucoma. In an embodiment, the disease condition can
be Alzheimer's disease. In some embodiments, the disease condition
can be a cognitive disorder.
BRIEF DESCRIPTION OF THE DRAWINGS
[0019] FIG. 1 is a graph that illustrates the results of an
amphetamine-induced hyperactivity study and illustrates distance
traveled as a function of tacrine dosage.
[0020] FIG. 2 is a graph that illustrates the results of an
amphetamine-induced hyperactivity study and illustrates percent
inhibition as a function of tacrine dosage.
[0021] FIG. 3 is a graph that illustrates the results of an
amphetamine-induced hyperactivity study and illustrates distance
traveled as a function of xanomeline dosage.
[0022] FIG. 4 is a graph that illustrates the results of an
amphetamine-induced hyperactivity study and illustrates percent
inhibition as a function of xanomeline dosage.
[0023] FIG. 5 is a bar graph that illustrates the results of a
novel object recognition study and illustrates the percentage of
time spent exploring a novel object as a function of tacrine
dosage.
[0024] FIG. 6 is a bar graph that illustrates the results of a
novel object recognition study and illustrates the percentage of
time spent exploring a novel object in relation to the drug or
combination of drugs administered.
[0025] FIG. 7 is a graph that illustrates the results of an
amphetamine-induced hyperactivity study and illustrates distance
traveled as a function of time.
[0026] FIG. 8 is a graph that illustrates the results of an
amphetamine-induced hyperactivity study and illustrates distance
traveled as a function of time.
[0027] FIG. 9 is a graph that illustrates the results of an
amphetamine-induced hyperactivity study M.sub.1 knock-out mice and
illustrates distance traveled as a function of pimavanserin
dosage.
[0028] FIG. 10 is a graph that illustrates the results of an
amphetamine-induced hyperactivity study in M.sub.1 knock-out mice
and illustrates percent inhibition as a function of pimavanserin
dosage.
DETAILED DESCRIPTION
[0029] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as is commonly understood by one
of ordinary skill in the art. All patents, applications, published
applications and other publications referenced herein are
incorporated by reference in their entirety.
[0030] It is understood that, in any compound described herein
having one or more chiral centers, if an absolute stereochemistry
is not expressly indicated, then each center may independently be
of R-configuration or S-configuration or a mixture thereof. Thus,
the compounds provided herein may be enatiomerically pure or be
stereoisomeric mixtures. In addition it is understood that, in any
compound described herein having one or more double bond(s)
generating geometrical isomers that can be defined as E or Z each
double bond may independently be E or Z a mixture thereof.
Likewise, all tautomeric forms are also intended to be
included.
[0031] An "agonist" is defined as a compound that increases the
basal activity of a receptor (i.e. signal transduction mediated by
the receptor).
[0032] An "inverse agonist" is defined as a compound that decreases
the basal activity of a receptor (i.e., signaling mediated by the
receptor). Such compounds are also known as negative antagonists.
An inverse agonist is a ligand for a receptor that causes the
receptor to adopt an inactive state relative to a basal state
occurring in the absence of any ligand. Thus, while an antagonist
can inhibit the activity of an agonist, an inverse agonist is a
ligand that can alter the conformation of the receptor in the
absence of an agonist. The concept of an inverse agonist has been
explored by Bond et al. in Nature 374:272 (1995). More
specifically, Bond et al. have proposed that ligand free
.beta..sub.2-adrenoceptor exists in an equilibrium between an
inactive conformation and a spontaneously active conformation.
Agonists are proposed to stabilize the receptor in an active
conformation. Conversely, inverse agonists are believed to
stabilize an inactive receptor conformation. Thus, while an
antagonist manifests its activity by virtue of inhibiting an
agonist, an inverse agonist can additionally manifest its activity
in the absence of an agonist by inhibiting the spontaneous
conversion of an unliganded receptor to an active conformation.
[0033] As used herein, "antagonist" refers to a compound that
competes with an agonist or inverse agonist for binding to a
receptor, thereby blocking the action of an agonist or inverse
agonist on the receptor. An antagonist attenuates the action of an
agonist on a receptor. However, an antagonist (also known as a
"neutral agonist") has no effect on constitutive receptor activity.
An antagonist may bind reversibly or irreversibly, and may reduce
the activity of the receptor until the antagonist is metabolized or
dissociates or is otherwise removed by a physical or biological
process.
[0034] The terms "pure," "purified," "substantially purified," and
"isolated" as used herein refer to the compound of the embodiment
being free of other, dissimilar compounds with which the compound,
if found in its natural state, would be associated in its natural
state. In some embodiments described as "pure," "purified,"
"substantially purified," or "isolated" herein, the compound may
comprise at least 75%, 80%, 85%, 90%, 95%, 99% of the mass, by
weight, of a given sample.
[0035] Pimavanserin, which is also known as
N-(1-methylpiperidin-4-yl)-N-(4-fluorophenylmethyl)-N'-(4-(2-methylpropyl-
oxy)phenylmethyl)carbamide,
N-[(4-fluorophenyl)methyl]-N-(1-methyl-4-piperidinyl)-N'-[[4-(2-methylpro-
poxy)phenyl]methyl]-urea,
1-(4-fluorobenzyl)-1-(1-methylpiperidin-4-yl)-3-[4-(2-methylpropoxy)benzy-
l]urea, or ACP-103 has the structure of Formula (I):
##STR00001##
[0036] Pimavanserin exhibits activity at serotonin receptors, and
acts as an inverse agonist of the 5-HT2A receptor. Experiments
performed on cells transiently expressing the human phenotype of
the 5-HT2A receptor have shown that pimavanserin attenuates the
signaling of such receptors in the absence of additional ligands
acting upon the receptor. Pimavanserin has thus been found to
possess inverse agonist activity at the 5-HT2A receptor and is able
to attenuate the basal, non-agonist-stimulated, constitutive
signaling responses that this receptor displays. The observation
that pimavanserin is an inverse agonist of the 5-HT2A receptor also
indicates that it has the ability to antagonize the activation of
5-HT2A receptors that is mediated by endogenous agonists or
exogenous synthetic agonist ligands. Pimavanserin exhibits high
affinity for the 5-HT2A receptor with a pK.sub.i>9. In vivo
human and non-human animal studies have further shown that
pimavanscrin exhibits antipsychotic, anti-dyskinesia, and
anti-insomnia activity. Such properties of pimavanserin are
described in U.S. Patent Publication No. 2004-0213816, filed Jan.
15, 2004 and entitled, "SELECTIVE SEROTONIN 2A/2C RECEPTOR INVERSE
AGONISTS AS THERAPEUTICS FOR NEURODEGENERATIVE DISEASES," which is
incorporated herein by reference in its entirety, including any
drawings.
[0037] Pimavanserin exhibits selective activity at the 5-HT2A
receptor. Specifically, pimavanserin lacks functional activity
(pEC.sub.50 or pK.sub.i<6) at 31 of the 36 human monoaminergic
receptors including 5-HT1A, 5-HT1B, 5-HT1D, 5-HT1E, 5-HT1F, 5-HT2B,
5-HT3, 5-HT4, 5-HT6A, 5-HT7A, adrenergic-.alpha.1A,
adrenergic-.alpha.1B, adrenergic-.alpha.1D, adrenergic-.alpha.2A,
adrenergic-.alpha.2B, adrenergic-.beta.2, dopamine-D1, dopamine-D2,
dopamine-D3, dopamine-D4, histamine-H1, histamine-H2, and
histamine-H3. Thus, pimavanserin provides high affinity at 5-HT2A
receptors with little to no affinity to most other monoaminergic
receptors. In one embodiment, pimavanserin exhibits a pK.sub.i of
less than 6 to dopamine receptors including the D2 and D4
receptors.
[0038] In addition, pimavanserin exhibits high stability, good oral
bioavailability, and a long half-life. Specifically, pimavanserin
exhibited a slow clearance rate from in vitro human microsomes
(<10 .mu.Lminmg) and a half-life of approximately 55 hours upon
oral administration to humans.
[0039] Various forms of pimavanserin can be used in the composition
with the agent that ameliorates one or more cholinergic
abnormalities. For example, a number of salts and crystalline forms
of pimavanserin can be used. Exemplary salts include the tartrate,
hemi-tartrate, citrate, fumarate, maleate, malate, phosphate,
succinate, sulphate, and edisylate (ethanedisulfonate) salts.
Pimavanserin salts including the aforementioned ions, among others,
are described in U.S. Patent Publication No. 2006-0111399, filed
Sep. 26, 2005 and entitled "SALTS OF
N-(4-FLUOROBENZYL)-N-(1-METHYLPIPERIDIN-4-YL)-N'-(4-(2-METHYLPROPYLOXY)PH-
ENYLMETHYL)CARBAMIDE AND THEIR PREPARATION," which is incorporated
herein by reference in its entirety. Two crystalline forms of the
tartrate salt are referred to as crystalline Form A and Form C,
respectively, and are described in U.S. Patent Publication No.
2006-0106063, filed Sep. 26, 2005 and entitled "SYNTHESIS OF
N-(4-FLUOROBENZYL)-N-(1-METHYLPIPERIDIN-4-YL)-N'-(4-(2-METHYLPROPYLOXY)PH-
ENYLMETHYL)CARBAMIDE AND ITS TARTRATE SALT AND CRYSTALLINE FORMS,"
which is incorporated herein by reference in its entirety.
Pimavanserin (including, for example, the tartrate salt) may be
formulated into tablets, such as is described in more detail in
U.S. Patent Publication Nos. 2007-0260064, filed May 15, 2007 and
2007-0264330, filed May 15, 2007, each entitled "PHARMACEUTICAL
FORMULATIONS OF PIMAVANSERIN," which are incorporated herein by
reference in their entireties.
[0040] Similarly, isolated, substantially pure metabolites of
pimavanserin can also be used. Suitable metabolites that can be
used have the chemical structures of Formulae (II), (I), (IV), (V)
and (VI) shown below.
##STR00002##
[0041] Compounds of Formulae (II), (III), (IV), (V) and (VI) as
described herein may be prepared in various ways. General synthetic
routes to the compounds of Formulae (II), (III), (IV), (V) and (VI)
are shown in Schemes A-E. The routes shown are illustrative only
and are not intended, nor are they to be construed, to limit the
scope of this invention in any manner whatsoever. Those skilled in
the art will be able to recognize modifications of the disclosed
synthesis and to devise alternate routes based on the disclosures
herein; all such modifications and alternate routes are within the
scope of this invention.
##STR00003##
[0042] Scheme A shows a general reaction scheme for forming the
compound of Formula (II). As shown in Scheme A, the secondary amine
and isocyanate can be combined to produce the 4-methoxybenzyl
derivative of the compound of Formula (II). The methoxy group can
be converted to a hydroxy group using methods known to those
skilled in the art, for example, using a boron trihalide to form
the compound of Formula (II).
##STR00004##
[0043] An exemplary method for synthesizing the compound of Formula
(III) is shown in Scheme B. The protected 4-piperidoinone and
4-fluorobenzylamine can undergo reductive amination to form
N-(4-fluorobenzyl)-4-amino 1-trifluoroacetylpiperidine. The
resulting secondary amine can then be reacted with the appropriate
isocyanate to form the nitrogen-protected carbamide. The acyl
protecting group can be cleaved off using an alkali metal salt such
as potassium carbonate to form the compound of Formula (III).
##STR00005##
[0044] One method for synthesizing the compound of Formula (IV) is
shown in Scheme C. The compound of Formula (II) can be reacted with
isobutylene oxide to form the compound of Formula (IV) via a
nucleophilic ring opening of the epoxide.
##STR00006##
[0045] Scheme D shows a general reaction scheme for forming the
compound of Formula (V). As shown in Scheme D, the compound of
Formula (II) can be reacted with a halohydrin to form the compound
of Formula (V). All the compounds described herein can be purified
using methods known to those skilled in art.
##STR00007##
[0046] One example of a method for synthesizing a compound of
Formula (VI) is shown in Scheme E. As shown in Scheme E,
N-(1-methylpiperidin-4-yl)-N-(4-fluorophenylmethyl)-N'-(4-(2-methylpropyl-
oxy)phenylmethyl) carbamide can be oxidized with a suitable
oxidizing agent to form a compound of Formula (VI). Suitable
oxidizing agents are known to those skilled in the art. One example
of a suitable oxidizing agent is meta-chloroperbenzoic acid. All
the compounds described herein can be purified using methods known
to those skilled in art.
[0047] When pimavanserin is used herein, it is understood that
salts, hydrates, polymorphs and isolated, substantially pure
metabolites thereof, either individually or in combination could be
used in place of pimavanserin. In an embodiment, the form of
pimavanserin that can be used is its tartrate salt.
[0048] Some embodiments disclosed herein relate to the
co-administration of pimavanserin with an agent that ameliorates
one or more cholinergic abnormalities. By "co-administration," or,
administration in "combination," it is meant that the two or more
agents may be found in the subject's bloodstream at the same time,
regardless of when or how they are actually administered. In some
embodiments, pimavanserin can be administered at approximately the
same time with the agent ameliorates one or more cholinergic
abnormalities. As used herein, at approximately the same time means
at substantially the same time or without a measure amount of time
between. For example, simultaneous administration can be
accomplished by combining pimavanserin and the agent that
ameliorates one or more cholinergic abnormalities in a single
dosage form. In another embodiment, pimavanserin and the agent that
ameliorates one or more cholinergic abnormalities can be
administered sequentially. For example, pimavanserin and the agent
that ameliorates one or more cholinergic abnormalities can each be
formulated as a separate dose. In an embodiment, pimavanserin can
be administered before the agent that ameliorates one or more
cholinergic abnormalities. In another embodiment, pimavanserin can
be administered after the agent that ameliorates one or more
cholinergic abnormalities. In another embodiment, a measurable
amount of time elapses between each administration. In one
embodiment, pimavanserin and the agent that ameliorates one or more
cholinergic abnormalities can be administered through the same
route, such as orally. In another embodiment, pimavanserin and the
agent that ameliorates one or more cholinergic abnormalities are
administered through different routes, such as one being
administered orally and another being administered intravenously
(i.v.). In one embodiment, the pharmacokinetics of pimavanserin and
the agent that ameliorates one or more cholinergic abnormalities
are substantially the same.
[0049] It has been surprisingly discovered that co-administration
of pimavanserin with an agent that ameliorates one or more
cholinergic abnormalities can synergistically affect the efficacy
of the agent. Some embodiments disclosed herein relate to a
composition that can include pimavanserin and an agent that
ameliorates one or more cholinergic abnormalities. Some embodiments
disclosed herein relate to a method for treating or ameliorating a
disease condition by administering pimavanserin in combination with
an agent that ameliorates one or more cholinergic abnormalities to
attain a synergistic effect. In an embodiment, the agent can
promote the activity of an M.sub.1 muscarinic receptor.
[0050] As previously mentioned, administration of pimavanserin in
combination with an agent that ameliorates one or more cholinergic
abnormalities has been found to synergistically enhance the
efficacy of the agent. Agents that ameliorate cholinergic
abnormalities may act through a variety of mechanisms, including
but not limited to, increasing acetylcholine concentration,
increasing the activity of the muscarinic receptors (e.g., M1
receptor), rectifying alternations in choline transport, rectifying
impaired acetylcholine release, addressing deficits in the
nicotinic and muscarinic receptor expression, rectifying
dysfunctional neurotrophin support, and/or rectifying deficits in
axonal support. In some embodiments, the agent that ameliorates one
or more cholinergic abnormalities can be selected from a
cholinesterase inhibitor, a muscarinic agonist, a glutamatergic
antagonist, a cholinergic agonist, a carnitine acetyltransferase
stimulant, an acetylcholine release stimulant, and a choline uptake
stimulant.
[0051] In some embodiments, the agent that ameliorates one or more
cholinergic abnormalities can be a cholinesterase inhibitor. In an
embodiment, the cholinesterase inhibitor can be selected from an
acetylcholinesterase inhibitor and a butyrylcholinesterase
inhibitor. Exemplary cholinesterase inhibitors include, but are not
limited to, organophosphates such as metrifonate, echothiophate and
diisopropyl fluorophosphate; carbamates such as physostigmine,
neostigmine, pyridostigmine, ambenonium, demarcarium, rivastigmine,
aldicarb, bendiocarb, bufencarb, carbaryl, carbendazim,
carbetamide, carbofuran, chlorbufam, chloropropham, ethiofencarb,
formetanate, methiocarb, methomyl, oxamyl, phenmedipham, pinmicarb,
pirimicarb, propamocarb, propham, and propoxur; phenanthrine
derivatives such as galantamine; piperidines such as donepezil;
tacrine; edrophonium; phenothiazines; dimebon
(3,6-dimethyl-9-(2-methyl-pyridyl-5)-ethyl-1,2,3,4-tetrahydro-.ga-
mma.-carboline dihydrochloride); Huperzine A; T-82
((2-[2-(1-benzylpiperidin-4-yl)ethyl]-2,3-dihydro-9-methoxy-1H-pyrrolo[3,-
4-b]quinolin-1-one hemifumarate)); TAK-147 (zanapezil); phenserine;
quilostigmine; ganstigmine; and butylcholinesterase inhibitors such
as butyrophenones, imipramines, tropates, phencyclidines,
curariforms, ethephon, ethopropazine, iso-OMPA,
tetrahydrofurobenzofuran cymserine, N.sup.1phenethyl-norcymserinc,
N.sup.8-benzylnorcymserine, N.sup.1,N.sup.8-bisnorcymserine,
N.sup.1--N.sup.8-bisbenzylnorphysostigmine,
N.sup.1,N.sup.8-bisbenzylnorphenserine and
N.sup.1,N.sup.8-bisbenzylnorcymserine.
[0052] In other embodiments, the agent that ameliorates one or more
cholinergic abnormalities can be a muscarinic agonist. Exemplary
muscarinic receptor agonists include, but are not limited to,
xanomeline, carbamylcholine, oxotremorine, methacholine,
bethanechol, cevimeline (AF102B), AF150(S), AF267B, aceclidine,
arecoline, milameline, talsaclidine, pilocarpine and
(S)-2-ethyl-8-methyl-1-thia-4,8-diaza-spiro[4.5]decan-3-one (Torrey
Pines NGX267). In one embodiment, the muscarinic agonist can be
xanomeline.
[0053] In still other embodiments, the agent that ameliorates one
or more cholinergic abnormalities can be a glutamatergic
antagonist. Exemplary glutamatergic antagonists include, but are
not limited to, memantine, amantadine, dextromethorphan,
dextrorphan, ibogaine, ketamine, tramadol, and methadone.
[0054] In some embodiments, the agent that ameliorates one or more
cholinergic abnormalities can be a cholinergic agonist. Exemplary
cholinergic agonists include, but are not limited to, pramiracetam,
piracetam, oxiracetam, choline-L-alfoscerate, nebracetam,
besipirdine, and taltirelin.
[0055] In other embodiments, the agent that ameliorates one or more
cholinergic abnormalities can be a carnitine acetyltransferase
stimulant. Exemplary carnitine acetyltransferase stimulants
include, but are not limited to, levocarnitine, ST-200
(acetyl-1-carnitine), and nefiracetam.
[0056] In still other embodiments, the agent that ameliorates one
or more cholinergic abnormalities can be an acetylcholine release
stimulant. Exemplary acetylcholine release stimulants include, but
are not limited to, SIB-1553A
((+/-)-4-[[2-(1-methyl-2-pyrrolidinyl)ethyl]thio]phenol
hydrochloride) and T-588
((1R)-1-benzo[b]thiophen-5-yl-2-[2-(diethylamino) ethoxy]ethan-1-ol
hydrochloride).
[0057] In even other embodiments, the agent that ameliorates one or
more cholinergic abnormalities can be a choline uptake stimulant.
In an embodiment, the choline uptake stimulant can be MKC-231
(2-(2-oxopyrrolidin-1-yl)-N-(2,3-dimethyl-5,6,7,8-tetrahydrofuro[2,3-b]qu-
inolin-4-yl)acetoamide).
[0058] Other embodiments disclosed herein relate to a
pharmaceutical composition that can include pimavanserin, an agent
that ameliorates one or more cholinergic abnormalities, and a
pharmaceutically acceptable salt, carrier, diluent, and/or
excipient.
[0059] As used herein, "pharmaceutically acceptable salt" refers to
a salt of a compound that does not abrogate the biological activity
and properties of the compound. Pharmaceutical salts can be
obtained by reaction of a compound disclosed herein with an acid or
base. Base-formed salts include, without limitation, ammonium salt
(NH.sub.4.sup.+); alkali metal, such as, without limitation, sodium
or potassium, salts; alkaline earth, such as, without limitation,
calcium or magnesium, salts; salts of organic bases such as,
without limitation, dicyclohexylamine, N-methyl-D-glucamine,
tris(hydroxymethyl)methylamine; and salts with the amino group of
amino acids such as, without limitation, arginine and lysine.
Useful acid-based salts include, without limitation,
hydrochlorides, hydrobromides, sulfates, nitrates, phosphates,
methanesulfonates, ethanesulfonates, p-toluenesulfonates and
salicylates.
[0060] As used herein, a "carrier" refers to a compound that
facilitates the incorporation of a compound into cells or tissues.
For example, without limitation, dimethyl sulfoxide (DMSO) is a
commonly utilized carrier that facilitates the uptake of many
organic compounds into cells or tissues of a subject.
[0061] As used herein, a "diluent" refers to an ingredient in a
composition that lacks pharmacological activity but may be
pharmaceutically necessary or desirable. For example, a diluent may
be used to increase the bulk of a potent drug whose mass is too
small for manufacture or administration. It may also be a liquid
for the dissolution of a drug to be administered by injection,
ingestion or inhalation. A common form of diluent in the art is a
buffered aqueous solution such as, without limitation, phosphate
buffered saline that mimics the composition of human blood.
[0062] As used herein, an "excipient" refers to an inert substance
that is added to a composition to provide, without limitation,
bulk, consistency, stability, binding ability, lubrication,
disintegrating ability etc., to the composition. A "diluent" is a
type of excipient.
[0063] The pharmaceutical compositions disclosed herein may be
manufactured in a manner that is itself known, e.g., by means of
conventional mixing, dissolving, granulating, dragee-making,
levigating, emulsifying, encapsulating, entrapping or tableting
processes. Proper formulation is dependent upon the route of
administration chosen. Techniques for formulation of the
compositions described herein are known to those skilled in the
art. The active ingredients are contained in an amount effective to
achieve its intended purpose. Many of the compounds used in the
pharmaceutical combinations disclosed herein may be provided as
salts with pharmaceutically compatible counterions.
[0064] Some embodiments disclosed herein relate to a method for
ameliorating or treating a disease condition characterized by one
or more cholinergic abnormalities that can include administering a
therapeutically effective amount of pimavanserin, and a
therapeutically effective amount an agent that ameliorates one or
more cholinergic abnormalities to a subject suffering from the
disease condition characterized by one or more cholinergic
abnormalities. Cholinergic abnormalities include, but are not
limited to the following: alternations in choline transport,
impaired acetylcholine release, deficits in the expression of
nicotinic and muscarinic receptors, dysfunctional neurotrophin
support and deficits in axonal transport. The aforementioned are
classified as abnormalities when compared to a healthy subject.
Other embodiments disclosed herein relate to a method for
ameliorating or treating a disease condition characterized by one
or more cholinergic abnormalities that can include administering a
therapeutically effective amount of pimavanserin, and a
therapeutically effective amount of an agent that ameliorates one
or more cholinergic abnormalities to a subject suffering from the
disease condition characterized by one or more cholinergic
abnormalities. In some embodiments, the agent that ameliorates one
or more cholinergic abnormalities can be selected from a
cholinesterase inhibitor, a muscarinic agonist, and a glutamatergic
agonist. In an embodiment, the agent promotes the activity of an
M.sub.1 muscarinic receptor.
[0065] Conditions suitable for treatment by a composition disclosed
herein include, but are not limited to, cognitive impairment,
forgetfulness, confusion, memory loss, an attention deficit
disorder, deficits in visual perception, depression, pain, sleep
disorders, psychosis, increased intraocular pressure,
neurodegenerative diseases, Alzheimer's disease, Parkinson's
disease, schizophrenia, Huntington's chorea, Friederich's ataxia,
Gilles de la Tourette's Syndrome, Down Syndrome, Pick disease,
dementia, clinical depression, age-related cognitive decline,
attention-deficit disorder, sudden infant death syndrome, glaucoma,
mania, bipolar disorder, unipolar disorder, schizoaffective
disorder, schizophreniform disorder and anxiety. It should be noted
that other non-schizophrenic causes of psychosis, including
drug-induced and treatment-induced, those associated with dementia
and other neurodegenerative disorders (such as Huntington's and
Alzheimer's) are also suitable.
[0066] In some embodiments, pimavanserin and an agent that
ameliorates one or more cholinergic abnormalities can be
administered in combination to treat, ameliorate, or prevent a
neuropsychiatric disorder, including but not limited to
schizophrenia, schizoaffective disorders, mania, depression
(including dysthymia, treatment-resistant depression, and
depression associated with psychosis), cognitive disorders,
aggressiveness (including impulsive aggression), a panic attack, an
obsessive compulsive disorder, borderline personality disorder,
borderline disorder, multiplex developmental disorder (MDD), a
behavioral disorder (including behavioral disorders associated with
age-related dementia), psychosis (including psychosis associated
with dementia, induced by treatment, such as treatment of
Parkinson's disease, or associated with post traumatic stress
disorder), suicidal tendency, bipolar disorder, sleep disorder
(including sleep maintenance insomnia, chronic insomnia, transient
insomnia, and periodic limb movements during sleep (PLMS)),
addiction (including drug or alcohol addiction, opioid addiction,
and nicotine addiction), attention deficit hyperactivity disorder
(ADHD), post traumatic stress disorder (PTSD), Tourette's syndrome,
anxiety (including general anxiety disorder (GAD)), autism, Down's
syndrome, a learning disorder, a psychosomatic disorder, alcohol
withdrawal, epilepsy, pain (including chronic pain, neuropathic
pain, inflammatory pain, diabetic peripheral neuropathy,
fibromyalgia, postherpetic neuralgia, and reflex sympathetic
dystrophy), a disorder associated with hypoglutamatergia (including
schizophrenia, childhood autism, and dementia), and serotonin
syndrome.
[0067] In some embodiments, pimavanserin can be administered in
combination with an agent that ameliorates one or more cholinergic
abnormalities to treat, ameliorate, or prevent a neurodegenerative
disorder. Exemplary neurodegenerative disorders include but are not
limited to, Alzheimer's disease (including psychosis and/or
dementia associated with Alzheimer's disease), Parkinson's disease,
Huntington's chorea, sphinocerebellar atrophy, frontotemporal
dementia, supranuclear palsy, or Lewy body dementia. In one
embodiment, the co-administration of pimavanserin and an agent that
ameliorates one or more cholinergic abnormalities can be used to
treat Alzheimer's disease with psychosis. In some embodiments,
symptoms associated with Alzheimer's disease that can be treated,
include, but are not limited to, agitation, aggression, delusions,
hallucinations, depression, insomnia, and anxiety.
[0068] In some embodiments, pimavanserin can be administered in
combination with an agent that ameliorates one or more cholinergic
abnormalities to treat, ameliorate, or prevent an extrapyramidal
disorder. Examples of extrapyramidal disorders include, but are not
limited to, dyskinesias (such as induced by treatment of
Parkinson's disease), bradykinesia, rigidity, psychomotor slowing,
tics, akathisia (such as induced by a neuroleptic or SSRI agent),
Friedrich's ataxia, Machado-Joseph's disease, dystonia, tremor,
restless legs syndrome, and myoclonus.
[0069] In some embodiments, pimavanserin can be administered in
combination with an agent that ameliorates one or more cholinergic
abnormalities to treat, ameliorate, or prevent psychosis.
Functional causes of the psychosis may include schizophrenia; a
bipolar disorder; severe clinical depression; severe psychosocial
stress; sleep deprivation; a neurological disorder including a
brain tumor; dementia with Lewy bodies; multiple sclerosis;
sarcoidosis; an electrolyte disorder including hypocalcemia,
hypernatremia, hyonatremia, hyopkalemia, hypomagnesemia,
hypermagnesania, hypercalcemia, hypophosphatemia, and hypoglycemia;
lupus; AIDS; leprosy; malaria; flu; mumps; psychoactive drug
intoxication; withdrawal including alcohol, barbiturates,
benzodiazepines, anticholinergics, atropine, scopolamine, Jimson
weed, antihistamines, cocaine, amphetamines, and hallucinogens such
as cannabis, LSD, psilocybin, mescaline, MDMA, and PCP. Psychosis
may include symptoms such as delusions, hallucinations,
disorganized speech, disorganized behavior, gross distortion of
reality, impaired mental capacity, impaired affective response,
fluctuating level of consciousness, poor motor co-ordination,
inability to perform simple mental tasks, disorientation as to
person, place or time, confusion, and/or memory impairment. In one
embodiment, the subject can be experiencing acute exacerbation of
psychosis. In some embodiments, a combination described herein can
be used to treat schizophrenia. In an embodiment, the psychosis can
be associated with schizophrenia. In one embodiment, the subject
has exhibited a prior response, or is currently exhibiting a
response, to antipsychotic therapy. In one embodiment, the subject
exhibits a moderate degree of psychopathology.
[0070] In one embodiment, the co-administration of pimavanserin
with an agent that ameliorates one or more cholinergic
abnormalities can be used to treat, ameliorate, or prevent
depression.
[0071] In some embodiments, pimavanserin potentiates the ability of
an agent that ameliorates one or more cholinergic abnormalities to
have its therapeutic effect. For example, in some embodiments,
pimavanserin potentiates the ability of a cholinesterase inhibitor,
a muscarinic agonist, and/or a glutamatergic antagonist to have its
therapeutic effect. In some embodiments, pimavanserin potentiates
the ability of a cholinesterase inhibitor, a muscarinic agonist,
and/or a glutamatergic antagonist to treat psychosis and/or improve
cognition.
[0072] In some embodiments, the co-administration of pimavanserin
with an agent that ameliorates one or more cholinergic
abnormalities can be used to treat, ameliorate or prevent
chemotherapy-induced emesis, frailty, on/off phenomena,
non-insulin-dependent diabetes mellitus, metabolic syndrome, an
autoimmune disorder (including lupus and multiple sclerosis),
sepsis, increased intraocular pressure, glaucoma, a retinal disease
(including age related macular degeneration), Charles Bonnet
syndrome, substance abuse, sleep apnea, pancreatis, anorexia,
bulimia, a disorder associated with alcoholism, cerebral vascular
accidents, amyotrophic lateral sclerosis, AIDS related dementia,
traumatic brain or spinal injury, tinnitus, menopausal symptoms
(such as hot flashes), sexual dysfunction (including female sexual
dysfunction, female sexual arousal dysfunction, hypoactive sexual
desire disorder, decreased libido, pain, aversion, female orgasmic
disorder, and ejaculatory problems), low male fertility, low sperm
motility, hair loss or thinning, incontinence, hemorrhoids,
migraine, hypertension, thrombosis (including thrombosis associated
with myocardial infarction, stroke, idiopathic thrombocytopenic
purpura, thrombotic thrombocytopenic purpura, and peripheral
vascular disease), abnormal hormonal activity (such as abnormal
levels of ACTH, corticosterone, rennin, or prolactin), a hormonal
disorder (including Cushing's disease, Addison's disease, and
hyperprolactinemia), a pituitary tumor (including a prolactinoma),
a side effect associated with a pituitary tumor (including
hyperprolactinemia, infertility, changes in menstruation,
amenorrhea, galactorrhea, loss of libido, vaginal dryness,
osteoporosis, impotence, headache, blindness, and double vision),
vasospasm, ischemia, a cardiac arrhythmia, cardiac insufficiency,
asthma, emphysema, and/or an appetite disorder.
[0073] In some embodiments, pimavanserin can be used to treat,
ameliorate and/or prevent psychosis. In an embodiment, the
psychosis is Alzheimer's disease-induced psychosis.
[0074] As used herein, the terms "treating," "treatment,"
"therapeutic," or "therapy" do not necessarily mean total cure or
abolition of the disease or condition. Any alleviation of any
undesired signs or symptoms of a disease or condition, to any
extent can be considered treatment and/or therapy. Furthermore,
treatment may include acts that may worsen the subject's overall
feeling of well-being or appearance.
[0075] The term "therapeutically effective amount" is used to
indicate an amount of an active compound, or pharmaceutical agent,
that elicits the biological or medicinal response indicated. For
example, a therapeutically effective amount of compound can be the
amount need to prevent, alleviate or ameliorate symptoms of disease
or prolong the survival of the subject being treated This response
may occur in a tissue, system, animal or human and includes
alleviation of the symptoms of the disease being treated.
Determination of a therapeutically effective amount is well within
the capability of those skilled in the art, especially in light of
the detailed disclosure provided herein. The therapeutically
effective amount of the compounds disclosed herein required as a
dose will depend on the route of administration, the type of
animal, including human, being treated, and the physical
characteristics of the specific animal under consideration. The
dose can be tailored to achieve a desired effect, but will depend
on such factors as weight, diet, concurrent medication and other
factors which those skilled in the medical arts will recognize.
[0076] As used herein, a "subject" refers to an animal that is the
object of treatment, observation or experiment. "Animal" includes
cold- and warm-blooded vertebrates and invertebrates such as fish,
shellfish, reptiles and, in particular, mammals. "Mammal" includes,
without limitation, mice, rats, rabbits, guinea pigs, dogs, cats,
sheep, goats, cows, horses, primates, such as monkeys, chimpanzees,
and apes, and, in particular, humans.
[0077] The compositions described herein can be administered to a
human subject per se, or in pharmaceutical compositions where they
are mixed with other active ingredients, as in combination therapy,
or carriers, diluents, excipients or combinations thereof.
Techniques for administrating the compositions described herein are
known to those skilled in the art. Suitable routes of
administration may, for example, include oral, rectal, topical
transmucosal, or intestinal administration; parenteral delivery,
including intramuscular, subcutaneous, intravenous, intramedullary
injections, as well as intrathecal, direct intraventricular,
intraperitoneal, intranasal, intraocular injections or as an
aerosol inhalant.
[0078] One may also administer the composition in a local rather
than systemic manner, for example, via injection of the compound
directly into the infected area, often in a depot or sustained
release formulation.
[0079] The compositions may, if desired, be presented in a pack or
dispenser device which may contain one or more unit dosage forms
containing the active ingredient. The pack may for example comprise
metal or plastic foil, such as a blister pack. The pack or
dispenser device may be accompanied by instructions for
administration. The pack or dispenser may also be accompanied with
a notice associated with the container in form prescribed by a
governmental agency regulating the manufacture, use, or sale of
pharmaceuticals, which notice is reflective of approval by the
agency of the form of the drug for human or veterinary
administration. Such notice, for example, may be the labeling
approved by the U.S. Food and Drug Administration for prescription
drugs, or the approved product insert. Compositions that can
include a compound described herein formulated in a compatible
pharmaceutical carrier may also be prepared, placed in an
appropriate container, and labeled for treatment of an indicated
condition.
[0080] As will be readily apparent to one skilled in the art, the
useful in vivo dosage to be administered and the particular mode of
administration will vary depending upon the age, weight, the
severity of the affliction, and mammalian species treated, the
particular compounds employed, and the specific use for which these
compounds are employed. (See e.g., Fingi et al. 1975, in "The
Pharmacological Basis of Therapeutics", which is hereby
incorporated herein by reference in its entirety, with particular
reference to Ch. 1, p. 1). The determination of effective dosage
levels, that is the dosage levels necessary to achieve the desired
result, can be accomplished by one skilled in the art using routine
pharmacological methods. Typically, human clinical applications of
products are commenced at lower dosage levels, with dosage level
being increased until the desired effect is achieved.
Alternatively, acceptable in vitro studies can be used to establish
useful doses and routes of administration of the compositions
identified by the present methods using established pharmacological
methods. In cases of administration of a pharmaceutically
acceptable salt, dosages may be calculated as the free base.
[0081] Although the exact dosage will be determined on a
drug-by-drug basis, in most cases, some generalizations regarding
the dosage can be made. Typically, the dose range of the
composition administered to the subject can be from about 0.5 to
about 1000 mg/kg of the subject's body weight, or about 1 to about
500 mg/kg, or about 10 to about 500 mg/kg, or about 50 to about 100
mg/kg of the subject's body weight. Additionally, the daily dosage
regimen for an adult human subject may be, for example, an oral
dose of between about 0.1 mg and about 500 mg of each ingredient,
preferably between about 1 mg and about 250 mg, e.g. about 5 to
about 200 mg or an intravenous, subcutaneous, or intramuscular dose
of each ingredient between about 0.01 mg and about 100 mg,
preferably between about 0.1 mg and about 60 mg, e.g. about 1 to
about 40 mg of each ingredient of the pharmaceutical compositions
disclosed herein or a pharmaceutically acceptable salt thereof
calculated as the free base. The dosage may be a single one or a
series of two or more given in the course of one or more days, as
is needed by the subject. In some embodiments, the compounds will
be administered for a period of continuous therapy, for example for
a week or more, or for months or years.
[0082] In instances where human dosages for compounds have been
established for at least some condition, the composition will use
those same dosages, or dosages that are between about 0.1% and
about 500%, more preferably between about 25% and about 250% of the
established human dosage. Where no human dosage is established, as
will be the case for newly-discovered pharmaceutical compositions,
a suitable human dosage can be inferred from ED.sub.50 or ID.sub.50
values, or other appropriate values derived from in vitro or in
vivo studies, as qualified by toxicity studies and efficacy studies
in animals.
[0083] As will be understood by those of skill in the art, in
certain situations it may be necessary to administer the compounds
disclosed herein in amounts that exceed, or even far exceed, the
above-stated, preferred dosage range in order to effectively and
aggressively treat particularly aggressive diseases or
infections.
[0084] Dosage amount and interval may be adjusted individually to
provide plasma levels of the active moiety which are sufficient to
maintain the modulating effects, or minimal effective concentration
(MEC). The MEC will vary for each compound but can be estimated
from in vitro data. Dosages necessary to achieve the MEC will
depend on individual characteristics and route of administration.
However, HPLC assays or bioassays can be used to determine plasma
concentrations.
[0085] Dosage intervals can also be determined using MEC value.
Compositions should be administered using a regimen which maintains
plasma levels above the MEC for 10-90% of the time, preferably
between 30-90% and most preferably between 50-90/%. In cases of
local administration or selective uptake, the effective local
concentration of the drug may not be related to plasma
concentration.
[0086] It should be noted that the attending physician would know
how to and when to terminate, interrupt, or adjust administration
due to toxicity or organ dysfunctions. Conversely, the attending
physician would also know to adjust treatment to higher levels if
the clinical response were not adequate (precluding toxicity). The
magnitude of an administrated dose in the management of the
disorder of interest will vary with the severity of the condition
to be treated and to the route of administration. The severity of
the condition may, for example, be evaluated, in part, by standard
prognostic evaluation methods. Further, the dose and perhaps dose
frequency, will also vary according to the age, body weight, and
response of the individual subject. A program comparable to that
discussed above may be used in veterinary medicine.
[0087] In non-human animal studies, applications of potential
products are commenced at higher dosage levels, with dosage being
decreased until the desired effect is no longer achieved or adverse
side effects disappear. The dosage may range broadly, depending
upon the desired effects and the therapeutic indication.
Alternatively dosages may be based and calculated upon the surface
area of the subject, as understood by those of skill in the
art.
[0088] Compounds disclosed herein can be evaluated for efficacy and
toxicity using known methods. For example, the toxicology of a
particular compound, or of a subset of the compounds, sharing
certain chemical moieties, may be established by determining in
vitro toxicity towards a cell line, such as a mammalian, and
preferably human, cell line. The results of such studies are often
predictive of toxicity in animals, such as mammals, or more
specifically, humans. Alternatively, the toxicity of particular
compounds in an animal model, such as mice, rats, rabbits, or
monkeys, may be determined using known methods. The efficacy of a
particular compound may be established using several recognized
methods, such as in vitro methods, animal models, or human clinical
trials. Recognized in vitro models exist for nearly every class of
condition, including but not limited to cancer, cardiovascular
disease, and various immune dysfunction. Similarly, acceptable
animal models may be used to establish efficacy of chemicals to
treat such conditions. When selecting a model to determine
efficacy, the skilled artisan can be guided by the state of the art
to choose an appropriate model, dose, and route of administration,
and regime. Of course, human clinical trials can also be used to
determine the efficacy of a compound in humans.
EXAMPLES
Chemistry
[0089] .sup.1H NMR spectra were recorded at 400 MHz on a Varian
Mercury-VX400 MHz spectrometer and chemical shifts are given in
.delta.-values [ppm] referenced to the residual solvent peak
chloroform (CDCl.sub.3) at 7.26 and methanol (CD.sub.3OD) at 3.31
ppm. Coupling constants, J, are reported in Hertz. Unless otherwise
stated, the NMR spectra of the compounds are described for their
free amine form. Column chromatography was carried out using silica
gel 60 (particle size 0.030-0.070 mm) from Merck. Materials and
solvents were of the highest grade available from commercial
sources and used without further purification. Reversed phase
C.sub.18 solid phase extraction cartridges (SPE) were DSC-18 2 g/12
mL columns from Discovery.TM. Solid Phase Extraction Products,
Supelco. Preparative HPLC was run on a Waters/Micromass HPLC/MS
using a diode array detector (190-450 nm) UV detector and Micromass
ZMD-mass-spectrometer with electrospray ionization. A YMC J'sphere
ODS H80 19.times.100 mm column was used. The mobile phase was 0.15%
TFA in water/acetonitrile with a gradient starting at 30%
acetonitrile, going to 100% acetonitrile over 13 min. The flow rate
was 17 mL/min.
[0090] HPLC/LCMS Method.
[0091] Samples were run on a Waters/Micromass HPLC/MS using a diode
array detector (190-450 nm) UV detector and Micromass
ZMD-mass-spectrometer with electrospray ionization. A Phenomenex
Luna C.sub.18 (2) 3 .mu.m, 75.times.4.6 mm column was used. The
mobile phase was 10 mM ammonium acetate in water/acetonitrile with
a gradient starting at 30% acetonitrile, going to 95% acetonitrile
over 12 min. The flow rate was 1.0 mL/min.
Preparation of Hydrochloride Salts
[0092] The tertiary amine products were dissolved in
dichloromethane, treated with an excess of 1M HCl in diethyl ether
and precipitated from n-heptane. The solvents were removed in vacuo
and after drying, the hydrochloride salts were obtained as
colorless solids in quantitative yield.
N-(4-Fluorobenzyl-N-(1-methylpiperidin-4-yl)-N'-(4-hydroxybenzyl)carbamide
hydrochoride
[0093] N-((4-Fluorophenyl)methyl)-4-amino-1-methylpiperidine was
prepared from 1-methylpiperidine-4-one (1.15 mL, 10 mmol), which
was dissolved in methanol (30 mL). 4-Fluorobenzylamine (1.25 mL, 10
mmol) was added and the pH was adjusted to 5 with acetic acid.
NaBH.sub.3CN (1.25 g, 20 mmol) was added and the reaction mixture
was stirred for 3 h, after which it was concentrated. 2M aqueous
NaOH (30 mL) was added and the mixture extracted with
dichloromethane (2.times.50 mL). The combined organic phases were
dried over Na.sub.2SO.sub.4, filtered and evaporated, and this
crude product was purified by Kugelrohr distillation to give the
desired product (1.1 g, 50%) as a clear oil.
[0094] N-(4-Fluorobenzyl)-4-amino-1-methylpiperidine (4.00 g, 18.0
mmol) was dissolved in dichloromethane (150 mL). 4-Methoxybenzyl
isocyanate (3.26 g, 20.0 mmol) in dichloromethane (50 mL) was added
dropwise and the mixture was stirred for 3 h at room temperature.
The crude mixture was concentrated and purified by flash
chromatography (0-10% methanol in dichloromethane) to give
N-((4-fluorophenyl)methyl)-N-(1-methylpiperidin-4-yl)-N'-((4-methoxypheny-
l)methyl)carbamide (4.91 g, 71%). This carbamide (4.91 g, 13.0
mmol) was dissolved in dry dichloromethane (50 mL). The solution
was cooled to 0.degree. C. and boron tribromide (1M in
dichloromethane, 39.0 mL, 39.0 mmol) was added dropwise, and the
mixture stirred for 20 h at room temperature. Water (50 mL) and
n-butanol (10 mL) were added and the phases separated. The aqueous
phase was extracted a second time with a mixture of dichloromethane
(50 mL) and n-butanol (10 mL). The combined organic phases were
evaporated and the resulting solid was purified by flash
chromatography (0-20% methanol in dichloromethane) to give a
semi-pure solid (3.17 g, 67%). An analytical amount (25 mg) of this
material was purified by preparative HPLC to give a colorless oil
(10 mg). LC-MS showed [M+H].sup.+=372 (characteristic fragment:
223). .sup.1H-NMR (CD.sub.3OD, 400 MHz, Free base): .delta.
7.25-6.62 (m, 8H), 4.46 (s, 2H), 4.22 (s, 2H), 4.15-4.06 (m, 1H),
2.89-2.82 (m, 2H), 2.23 (s, 3H), 2.14-2.05 (m, 2H), 1.74-1.61 (m,
4H).
[0095] The collected compound was converted into its hydrochloride
salt, which was obtained as a colorless solid.
N-(4-Fluorobenzyl-N'-(piperidin-4-yl-N'-(4-isobutoxybenzyl)carbamide
hydrochloride
[0096] 4-Piperidone hydrochloride monohydrate (4.0 g, 26.0 mmol)
was dissolved in dichloromethane (130 mL). After addition of
triethylamine (8.66 g, 85.8 mmol) the mixture was stirred for 10
min and then cooled to 0.degree. C. Trifluoroacetic anhydride (12.0
g, 57.2 mmol) was added dropwise under stirring. After 2 hours at
room temperature, the reaction was stopped by addition of water
(100 mL). The aqueous phase was extracted with dichloromethane
(2.times.100 mL). The combined organic phases were dried over
Na.sub.2SO.sub.4, filtered and concentrated to give
1-trifluoroacetyl-4-piperidone (5.07 g, 100%). 4-Fluorobenzylamine
(3.14 g, 25.9 mmol) and 1-trifluoroacetyl-4-piperidone (5.07 g,
25.9 mmol) were added to a solution of methanol adjusted to pH 5
with acetic acid (150 mL). The reaction mixture was stirred for 5
min and NaBH.sub.3CN (2.46 g, 38.9 mmol) was added slowly under
stirring. After 20 hours at room temperature the reaction was
concentrated. 2M aqueous NaOH (100 mL) was added and extracted with
dichloromethane (2.times.100 mL). The combined organic phases were
dried over Na.sub.2SO.sub.4, filtered and concentrated to give
N-(4-fluorobenzyl)-4-amino-1-trifluoroacetylpiperidine (2.91 g,
37%).
[0097] 4-Isobutoxybenzyl isocyanate was prepared from
4-isobutoxyphenylacetic acid (7.6 g, 36.5 mmol) (prepared according
to classical literature procedures from methyl
4-hydroxyphenylacetate by a Williamson ether synthesis with
isobutylbromide, followed by saponification of the ester. For an
alternative route see: Profft; Drux; J. Prakt. Chem. 1956, 4(3),
274-275), which is hereby incorporated by reference in its
entirety, and which was dissolved in THF (50 mL). Proton Sponge.TM.
(8.2 g, 38 mmol) was added, and the mixture was stirred for 15 min.
Diphenylphosphoryl azide (10.6 g, 38 mmol) was added dropwise and
the mixture was heated to reflux for 4 h. The mixture was cooled to
room temperature and placed in the freezer at -18.degree. C. for 20
h. The resulting white precipitate was vigorously stirred with
diethyl ether (250 mL) for 15 min and filtered. The filtrate was
evaporated to give the desired product, which was used without
further purification.
[0098] N-(4-fluorobenzyl)-4-amino-1-trifluoroacetylpiperidine (2.91
g, 9.6 mmol) was dissolved in dichloromethane (50 mL) and a
solution of 4-isobutoxybenzyl isocyanate (1.97 g, 9.6 mmol) in
dichloromethane (50 mL) was added. The reaction mixture was stirred
for 20 h and concentrated. The crude product was purified by flash
chromatography (0-5% methanol in dichloromethane) to give
N-(4-fluorobenzyl)-N-(1-trifluoroacetylpiperidin-4-yl)-N'-(4-isobutoxyben-
zyl)carbamide (3.90 g, 91%).
[0099] This carbamide (3.90 g, 8.7 mmol) was dissolved in methanol
(12 mL) and added to a 2M solution of potassium carbonate in
methanol (100 mL) under stirring. After 4 hours the methanol was
evaporated, and the aqueous phase was extracted with
dichloromethane (2.times.100 mL). The combined organic phases were
dried over Na.sub.2SO.sub.4, filtered and concentrated to give a
semi-pure solid (2.95 g, 85%). An analytical amount (200 mg) of
this crude product was purified by flash chromatography (10%
methanol in dichloromethane with 1% triethylamine) to give a
colorless solid (100 mg). LC-MS showed [M+H].sup.+=414
(characteristic fragment: 209). .sup.1H-NMR (CDCl.sub.3, 400 MHz,
Free base): .delta. 7.21-6.75 (m, 8H), 4.47-4.42 (m, 1H), 4.39 (t,
J=5.0 Hz, 1H), 4.35 (s, 2H), 4.27 (d, J=5.0 Hz, 2H), 3.68 (d, J=6.0
Hz, 2H), 3.13-3.06 (m, 211), 2.74-2.66 (m, 2H), 2.11-1.99 (m, 1H),
1.78-1.71 (m, 3H), 1.58-1.46 (m, 2H), 1.00 (d, J=6.0 Hz, 6H).
[0100] The collected compound was converted into its hydrochloride
salt, which was obtained as a colorless solid.
N-(4-Fluorobenzyl)-N-(1-methylpiperidin-4-yl-N'-[4-(2-hydroxy)isobutoxyben-
zyl]carbamide hydrochloride
[0101]
N-(4-Fluorobenzyl)-N-(1-methylpiperidin-4-yl)-N'-(4-hydroxybenzyl)
carbamide (375 mg, 1.0 mmol) was dissolved in DMF (15 mL). KOH (281
mg, 5.0 mmol) was added and the mixture was stirred 30 min at room
temperature. Isobutylene oxide (216 mg, 3.0 mmol) was added and the
mixture was warmed to 40.degree. C. for 20 h. Isobutylene oxide
(216 mg, 3.0 mmol) was added and the mixture was stirred at
40.degree. C. for another 20 h. Water (50 mL) was added and the
mixture was extracted with dichloromethane (2.times.60 mL). The
combined organic phases were dried over Na.sub.2SO.sub.4, filtered
and evaporated. The crude product was purified by flash
chromatography (5% methanol in dichloromethane) and subsequently by
passage over a C.sub.18-SPE cartridge, eluting with 30%
acetonitrile/water and 3.5 mM ammonium acetate buffer. The
acetonitrile was evaporated and the water phase was made alkaline
with aqueous ammonia. The product was extracted into
dichloromethane (2.times.100 mL), and the combined organic phases
were dried over Na.sub.2SO.sub.4, filtered and evaporated to give a
colorless oil (122 mg, 28%). LC-MS showed [M+H].sup.+=444
(characteristic fragment: 223). .sup.1H-NMR (CDCl.sub.3, 400 MHz,
Free base): .delta. 7.21-6.77 (m, 8H), 4.49-4.43 (t, J=5.5 Hz, 1H),
4.37-4.26 (m, 5H), 3.75 (s, 2H), 2.89-2.82 (m, 2H), 2.25 (s, 3H),
2.10-2.01 (m, 2H), 1.76-1.58 (m, 411), 1.33 (s, 6H).
[0102] The collected compound was converted into its hydrochloride
salt, which was obtained as a colorless solid.
N-(4-Fluorobenzyl)-N-(1-methyl-piperidin-4-yl)-N'-(4-(R)-[(3-hydroxy)-isob-
utoxy]benzyl)carbamide
[0103]
N-(4-Fluorobenzyl)-N-(1-methylpiperidin-4-yl)-N'-(4-hydroxybenzyl)c-
arbamide (75 mg, 0.20 mmol) was dissolved in DMF (3 mL). Potassium
hydroxide (56 mg, 1.00 mmol) was added and the mixture was stirred
15 minutes at room temperature. (R)-(-)-3-Bromo-2-methyl-1-propanol
(93 mg, 0.60 mmol) was added. The mixture was heated to 60.degree.
C. for 5 hours. The reaction mixture was cooled to room
temperature, added to dichloromethane (50 mL), and washed with 1M
potassium hydroxide (50 mL). The organic phase was dried over
Na.sub.2SO.sub.4, filtered and evaporated. The resulting oil was
purified by preparative HPLC to give
N-(4-Fluorobenzyl)-N-(1-methyl-piperidin-4-yl)-N'-(4-(R)-[(3-hydroxy)-iso-
butoxy]benzyl)carbamide as a colorless oil (5 mg, 6%). LC-MS showed
[M+H].sup.+=444 (characteristic fragment: 223). .sup.1H-NMR
(CDCl.sub.3, 400 MHz, free base): .delta. 7.20-6.78 (m, 8H), 4.47
(t, J=5 Hz, 1H), 4.35-4.29 (m, 3H), 4.27 (d, J=5.0 Hz, 2H),
3.92-3.89 (m, 2H), 3.68 (d, J=6.0 Hz, 2H), 2.89-2.82 (m, 2H), 2.25
(s, 3H), 2.22-2.13 (m, 1H), 2.10-2.02 (m, 2H), 1.83-1.76 (bs, 1H),
1.75-1.59 (m, 4H) 1.02 (d, J=6.0 Hz, 3H).
[0104] The collected compound was converted into its hydrochloride
salt, which was obtained as a colorless solid.
N-(4-Fluorobenzyl)-N-(1-methyl-1-oxopiperidin-4-yl)-N'-(4-isobutoxybenzyl)-
carbamide
[0105]
N-(4-Fluorobenzyl)-N-(1-methylpiperidin-4-yl)-N'-(4-isobutoxybenzyl-
)carbamide (100 mg, 0.234 mmol) was dissolved in dichloromethane
(10 mL). The solution was cooled to 0.sup..degree. C. and
meta-chloroperbenzoic acid (57-86%, 106 mg, 0.351 mmol) was added.
The reaction mixture was stirred for 20 h at room temperature,
after which it was washed with saturated aqueous NaHCO.sub.3 (10
mL). The organic phase was dried over Na.sub.2SO.sub.4, filtered
and evaporated. The resulting oil was purified by preparative HPLC
to give
N-(4-fluorobenzyl)-N-(1-methyl-1-oxopiperidin-4-yl)-N'-(4-isobutoxybenzyl-
)carbamide as a colorless oil (10 mg, 10%). LC-MS showed
[M+H].sup.+=444 (characteristic fragment: 239). .sup.1H-NMR
(CDCl.sub.3, 400 MHz): .delta. 7.20-6.76 (m, 8H), 4.63-4.53 (m,
2H), 4.43 (s, 2H), 4.24 (d, J=5.0 Hz, 2H), 3.66 (d, J=7.0 Hz, 2H),
3.31-3.24 (m, 4H), 3.19 (s, 3H), 2.62-2.51 (m, 2H), 2.10-1.99 (m,
1H), 1.69-1.62 (m, 2H), 1.00 (d, J=7.0 Hz, 6H).
In Vitro Determination of Receptor Activity
[0106] Receptor Selection and Amplification (R-SAT) Assays.
[0107] The functional receptor assay, Receptor Selection and
Amplification Technology (R-SAT), was used (with minor
modifications from the procedure described previously (Brann, M. R.
U.S. Pat. No. 5,707,798, 1998; Chem. Abstr. 1998, 128, 111548) to
screen compounds for efficacy at the 5-HT.sub.2A receptor. Briefly,
NIH3T3 cells were grown in 96 well tissue culture plates to 70-80%
confluence. Cells were transfected for 12-16 h with plasmid DNAs
using superfect (Qiagen Inc.) as per manufacturer's protocols.
R-SAT's were generally performed with 50 ng/well of receptor and 20
ng/well of .beta.-galactosidase plasmid DNA. All receptor and
G-protein constructs used were in the pSI mammalian expression
vector (Promega Inc) as described previously. The 5-HT.sub.2A or
5-HT.sub.2C receptor gene was amplified by nested PCR from brain
cDNA using the oligodeoxynucleotides based on the published
sequence (Saltzman et. al, Biochem. Biophys. Res. Comm. 1991, 181,
1469). For large-scale transfections, cells were transfected for
12-16 h, then trypsinized and frozen in DMSO. Frozen cells were
later thawed, plated at 10,000-40,000 cells per well of a 96 well
plate that contained drug. With both methods, cells were then grown
in a humidified atmosphere with 5% ambient CO.sub.2 for five days.
Media was then removed from the plates and marker gene activity was
measured by the addition of the .beta.-galactosidase substrate
o-nitrophenyl .beta.-D-galactopyranoside (ONPG, in PBS with 5%
NP-40). The resulting colorimetric reaction was measured in a
spectrophotometric plate reader (Titertek Inc.) at 420 nM. All data
were analyzed using the computer program XLFit (IDBSm). Efficacy is
the percent maximal repression compared to repression by a control
compound (ritanserin in the case of 5-HT.sub.2A). pIC.sub.50 is the
negative of the log(IC.sub.50), where IC.sub.50 is the calculated
concentration in Molar that produces 50% maximal repression.
Various metabolites of
N-(1-methylpiperidin-4-yl)-N-(4-fluorophenylmethyl)-N'-(4-(2-methylpropyl-
oxy)phenylmethyl) carbamide, including the compounds of Formulae
(II), (III), (IV), (V) and (VI) as well as other metabolites, were
assayed as described herein. The assayed metabolites demonstrated
varying activity levels with some of the metabolites exhibiting
levels too low for use as pharmaceuticals agents. Compounds of
Formulae (II), (III), (IV), (V) and (VI), however, demonstrated
high inverse agonist and antagonist activity as shown in the table
below. This data indicates compounds of Formulae (II), (III), (IV),
(V) and (VI) could be useful as pharmaceutical agents.
TABLE-US-00001 Inverse Agonist Antagonist pIC.sub.50 pKi Compound
5HT2A 5HT2C 5HT2A 5HT2C Formula (II) 7.5 -- 7.9 6.5 Formula (III)
8.6 -- 8.9 7 Formula (IV) 8.7 6.1 9 6.8 Formula (V) 8.5 6.7 -- 6.8
Formula (VI) 7.5 5.7 7.5 6.4
Example 1
Amphetamine-Induced Hyperactivity with Tacrine
[0108] This study was conducted in acrylic chambers (42 cm.times.42
cm.times.30 cm) equipped with 16 infrared photobeams along each
horizontal axis (front-to-back and side-to-side, from Accuscan
Instruments, Inc., Columbus, Ohio). Mice were initially
administered either vehicle (veh) or pimavanserin (0.03 mg/kg) 60
minutes prior to entering activity chambers. Vehicle or a drug
(either tacrine or xanomeline) was injected 30 minutes prior to
entering activity chambers. Amphetamine (3 mg/kg) was injected into
mice 15 minutes prior to entering motor activity chambers. Mice had
no prior exposure to the chambers and each dose combination was
tested in separate groups of mice.
[0109] Activity was measured as total distance traveled (DT) in
centimeters and was determined across a 15 minute session. Dose
response curves were constructed for the drug in the presence of
either vehicle or a fixed dose of pimavanserin. In order to
generate dose-response curves, raw DT data were converted to
percent inhibition, % MPI=((DT drug or drug combination-DT
amphetamine control)/(DT vehicle control-DT amphetamine
control))*100. A linear regression analysis was then conducted to
determine the theoretical dose that would produce a 50% reduction
of activity (ED.sub.50) and 95% confidence interval. The results of
these studies for tacrine and xanomeline are presented in FIGS.
1-4.
[0110] FIG. 1 illustrates the total distance traveled as a function
of the dosage of tacrine administered in mice given a vehicle
control or amphetamine. As shown in FIG. 1, as the dosage of
tacrine was increased, the total distance traveled decreased. This
shows the ability of tacrine to suppress amphetamine-induced
hyperactivity in mice. Also shown in FIG. 1 is that the
administration of tacrine in combination with pimavanserin further
reduced the total distance traveled of mice given amphetamine for a
particular dosage of tacrine. FIG. 2 illustrates the percent
inhibition of amphetamine-induced hyperactivity in mice as a
function of tacrine dosage. Increasing doses of tacrine resulted in
a greater inhibition of amphetamine-induced hyperactivity. By
comparison, administration of tacrine in combination with
pimavanserin further resulted in inhibition for a particular dosage
of tacrine. Thus, pimavanserin potentiates the ability of tacrine
to suppress amphetamine-induced hyperactivity in mice. FIGS. 3 and
4 illustrate the total distance traveled as a function of
xanomeline dosage and percent inhibition as a function of
xanomeline dosage, respectively in mice treated with or without
amphetamine. Similar results to tacrine as shown in FIGS. 1 and 2
were observed for xanomeline. These results demonstrated that
pimavanscrin enhanced the antipsychotic-like activity of
cholinesterase inhibitors and muscarinic receptor agonists.
Example 2
Novel Object Recognition
[0111] Subjects were male, C57 BK/6 mice purchased from Harlan
Laboratories, weighing 15-20 g upon arrival. Animals were housed 8
per cage with food and water available ad libitum. Animals were
housed on a 12 hr light cycle (lights on 6 am) for 4-7 days prior
to behavioral testing.
[0112] Novel object recognition (NOR) was conducted in a novel
environment consisting of a white plastic tub measuring
45.7.times.33.7.times.19 cm. Prior to each trial the bottom of the
tub was covered with a piece of plastic lined bench top paper.
There were two sets of identical objects chosen so that when given
an opportunity to explore, mice would evenly divide exploration
time between the objects. "A" objects were yellow, ceramic,
12-sided ramekins measuring 4 cm high.times.7 cm diameter. "B"
objects were 8.times.8.times.4 cm stainless steel, 4-sided
ramekins.
[0113] At the beginning of each test day, animals were placed in
groups of 6 into clean cages. Testing was conducted in three
phases: acclimation, sample and test.
[0114] In the acclimation phase, each group of six mice was placed
collectively into the NOR chamber and allowed to explore freely for
30 minutes. After acclimation animals were injected (dose and
pretreatment time varied by test drug) and placed back into the
cages to wait the pre-treatment interval.
[0115] During the sample phase, two identical objects ("A" or "B"
objects described above) were placed into the NOR chamber. Objects
were placed on diagonal corners of the long axis of the arena
approximately 5 cm from the walls, while subjects were placed into
one of the neutral corners (alternating across subjects). Each
mouse was placed into the NOR chamber one at a time and was allowed
to explore the chamber and the objects for 3 minutes. The time
spent exploring at each position was recorded. Directly sniffing or
touching the object was recorded as exploration. After 3 minutes,
each mouse was removed from the arena and placed back into its
cage.
[0116] The test phase was conducted one to two hours after the
sample phase. During the test, one familiar object (seen during
sample) and one novel object were placed into the chamber in the
same positions used during the sample phase, and each mouse was
allowed 3 minutes to explore. The test sessions were recorded on
video and scored by an observer blind to each subject's treatment
condition. Any time spent directly sniffing or touching an object
was counted as exploration. The object serving as the novel object
and the position where the novel object was placed were
counterbalanced across subjects. Prior to each trial (acclimation,
sample and test), all equipment was wiped with a Clorox wipe and
bench paper (cut to fit) was placed in the bottom of the
chamber.
[0117] In addition to time spent exploring each object
(T.sub.N=time spent exploring novel object, T.sub.F=time spent
exploring familiar object), two measures were determined for each
subject: exploration ratio (% of time spent exploring at novel
object) ER=T.sub.N*100/(T.sub.N+T.sub.F) and discrimination index
(preference for novel) DI=(T.sub.N-T.sub.F)/(T.sub.N+T.sub.F).
[0118] During the sample phase, it was expected that there would be
no preference for one position over another in any treatment
condition. During the test phase, vehicle treated animals were
expected to show a preference for the novel object after 1 hour
(indicating they recognize the familiar object), but were expected
to return to baseline exploration rates after 2 hours.
##STR00008##
[0119] The results of these studies using tacrine are depicted in
FIGS. 5 and 6. FIG. 5 illustrates the percent of time each subject
spent exploring a novel object as a function of the dosage of
tacrine received. As the dosage of tacrine increased, the percent
of time spent exploring a novel object also increased, Preference
for exploring the novel object suggests that the subject recognizes
and distinguishes that one object is novel, while the other object
is familiar. Thus, FIG. 5 demonstrates that tacrine improves the
cognition of a subject.
[0120] FIG. 6 compares novel object recognition data for four
groups of subjects. Each group was treated with vehicle,
pimavanserin, tacrine, or a combination of pimavanserin and
tacrine. As shown in FIG. 6, the percent of time spent exploring a
novel object was highest for the group treated with a combination
of pimavanserin and tacrine. Notably, this group performed better
than the groups that had received either pimavanserin or tacrine
individually. These findings demonstrate the ability of
pimavanserin to augment the pro-cognitive effects of tacrine.
Example 3
Hyperactivity Study with an Amyloid Protein
[0121] This study was conducted in acrylic chambers (42 cm.times.42
cm.times.30 cm) equipped with 16 infrared photobeams along each
horizontal axis (front-to-back and side-to-side, from Accuscan
Instruments, Inc. (Columbus, Ohio)). Subjects were mice receiving
ICV infusion of amyloid .beta. protein fragment (25-35) or ICV
infusion of saline (sham) for 7-10 days. This procedure has been
shown to result in the formation of amyloid plaques and impaired
cognition, thus simulating the effects of Alzheimer's disease.
[0122] An amphetamine-induced hyperactivity study was conducted on
the subjects. Subjects were injected with amphetamine (3 mg/kg) or
vehicle 15 minutes prior to entering motor activity chambers.
Activity was measured as a total distance traveled (DT) in
centimeters (cm) and was determined in 20 minute increments across
a 60 minute session. The results of this study are shown in FIG.
7.
[0123] FIG. 7 shows distance traveled as a function of time. Among
the subjects injected with saline, amyloid .beta. mice traveled the
same distance over a given amount of time as the control animals;
thus, these mice exhibited normal basal levels of locomotor
activity. Among the subjects injected with amphetamine, amyloid
.beta. mice traveled farther on average than control mice. The
findings demonstrate that amyloid .beta. mice show an augmented
response to amphetamine as compared to the control animals.
[0124] In a subsequent study, mice were pretreated with either
pimavanserin (0.3 mg/kg) or vehicle 60 min prior to behavioral
testing, and all subjects were subsequently injected with
amphetamine (3 mg/kg) 15 minutes prior to entering motor activity
chambers. Activity was measured as a total distance traveled (DT)
in centimeters (cm) and was determined in 20 minute increments
across a 60 minute session.
[0125] FIG. 8 illustrates distance traveled as a function of time
following amphetamine pretreatment. Among the subjects treated with
vehicle, the amyloid .beta. mice traveled farther than the control
mice. In contrast, among the subjects treated with pimavanserin,
subjects traveled similar distances regardless of whether they were
in the amyloid .beta. or control groups. Thus, in the control mice,
pimavanserin alone did not reduce the hyperactivity induced by
amphetamine. However, in a model of Alzheimer's disease model,
pimavanserin reversed the augmented locomotor response to
amphetamine. These results suggest that pimavanserin can be
effective in treating, ameliorating, or preventing psychosis
associated with Alzheimer's disease.
Example 4
Hyperactivity Study with M1 Knock-Out Mice
[0126] This study was conducted in acrylic chambers (42 cm.times.42
cm.times.30 cm) equipped with 16 infrared photobeams along each
horizontal axis (front-to-back and side-to-side, from Accuscan
Instruments, Inc. (Columbus, Ohio). Subjects were mice with a
deletion of the M.sub.1 muscarinic receptor (M.sub.1KO) or
wild-type (WT) controls. Vehicle (saline) or test compound
(pimavanserin, 0.03 or 0.3 mg/kg) was injected 45 min prior to
vehicle or amphetamine (3 mg/kg ip). Fifteen minutes later, mice
were placed in the locomotor activity chambers and their activity
was recorded. In order to generate dose-response curves, raw DT
data were converted to % MPI: % MPI==((DT drug-DT MK801
control)/(DT vehicle control-DT MK801control))*100. A linear
regression analysis was then conducted to determine the theoretical
dose that would produce a 50% reduction of activity (ED.sub.50) and
95% confidence interval. Mice had no prior exposure to the chambers
and each dose combination was tested in separate groups of mice.
The results of these studies are presented in FIGS. 9-10.
[0127] FIG. 9 illustrates the results of an amphetamine-induced
hyperactivity study, showing distance traveled as a function of
pimavanserin dosage. While not wanting to be bound by any
particular theory, psychosis in humans can be associated with
augmented dopamine in the striatum. Mice devoid of the muscarinic
M1 receptors are hyperactive which can be correlated to augmented
dopamine levels in the striatum. As shown in FIG. 9, administration
of pimavanserin alone had no significant effect on locomotor
activity compared with vehicle controls. However, upon
administration of amphetamine, M.sub.1KO mice had an augmented
locomotor response compared with WT controls. While pimavanserin
did not alter amphetamine-induced activity in WT subjects, it
reversed the augmented amphetamine response in M1KO subjects. Thus,
after pimavanserin treatment, amphetamine produced a similar
increase in activity in both WT and M1KO subjects.
[0128] FIG. 10 illustrates percent inhibition as a function of
pimavanserin dosage generated from the distance traveled data shown
in FIG. 9. As demonstrated by FIG. 10, pimavanserin increased the
inhibition of amphetamine-induced hyperactivity. The results in
FIGS. 9 and 10 indicate pimavanserin reduces augmented
amphetamine-induced hyperactivity in mice devoid of muscarinic M1
receptors. Since reversal of amphetamine-induced hyperactivity is
predictive of antipsychotic activity in humans, these data suggest
that pimavanserin can be effective in treating or ameliorating
psychosis associated with a deficiency in muscarinic receptor
activity such as Alzheimer's disease-induced psychosis.
[0129] Although the invention has been described with reference to
embodiments and examples, it should be understood that numerous and
various modifications can be made without departing from the spirit
of the invention. Accordingly, the invention is limited only by the
following claims.
* * * * *